L. L. Granke

Fungal Planet description sheets: 107–127

Novel species of microfungi described in the present study include the following from Australia: Phytophthora amnicola from still water, Gnomoniopsis smithogilvyi from Castanea sp., Pseudoplagiostoma corymbiae from Corymbia sp., Diaporthe eucalyptorum from Eucalyptus sp., Sporisorium andrewmitchellii from Enneapogon aff. lindleyanus, Myrmecridium banksiae from Banksia, and Pilidiella wangiensis from Eucalyptus sp. Several species are also described from South Africa, namely: Gondwanamyces wingfieldii from Protea caffra, Montagnula aloes from Aloe sp., Diaporthe canthii from Canthium inerne, Phyllosticta ericarum from Erica gracilis, Coleophoma proteae from Protea caffra, Toxicocladosporium strelitziae from Strelitzia reginae, and Devriesia agapanthi from Agapanthus africanus. Other species include Phytophthora asparagi from Asparagus officinalis (USA), and Diaporthe passiflorae from Passiflora edulis (South America). Furthermore, novel genera of coelomycetes include Chrysocrypta corymbiae from Corymbia sp. (Australia), Trinosporium guianense, isolated as a contaminant (French Guiana), and Xenosonderhenia syzygii, from Syzygium cordatum (South Africa). Pseudopenidiella piceae from Picea abies (Czech Republic), and Phaeocercospora colophospermi from Colophospermum mopane (South Africa) represent novel genera of hyphomycetes. Morphological and culture characteristics along with ITS DNA barcodes are provided for all taxa.

The cucurbit downy mildew pathogen Pseudoperonospora cubensis.

UNLABELLED Pseudoperonospora cubensis[(Berkeley & M. A. Curtis) Rostovzev], the causal agent of cucurbit downy mildew, is responsible for devastating losses worldwide of cucumber, cantaloupe, pumpkin, watermelon and squash. Although downy mildew has been a major issue in Europe since the mid-1980s, in the USA, downy mildew on cucumber has been successfully controlled for many years through host resistance. However, since the 2004 growing season, host resistance has been effective no longer and, as a result, the control of downy mildew on cucurbits now requires an intensive fungicide programme. Chemical control is not always feasible because of the high costs associated with fungicides and their application. Moreover, the presence of pathogen populations resistant to commonly used fungicides limits the long-term viability of chemical control. This review summarizes the current knowledge of taxonomy, disease development, virulence, pathogenicity and control of Ps. cubensis. In addition, topics for future research that aim to develop both short- and long-term control measures of cucurbit downy mildew are discussed. TAXONOMY Kingdom Straminipila; Phylum Oomycota; Class Oomycetes; Order Peronosporales; Family Peronosporaceae; Genus Pseudoperonospora; Species Pseudoperonospora cubensis. DISEASE SYMPTOMS Angular chlorotic lesions bound by leaf veins on the foliage of cucumber. Symptoms vary on different cucurbit species and varieties, specifically in terms of lesion development, shape and size. Infection of cucurbits by Ps. cubensis impacts fruit yield and overall plant health. INFECTION PROCESS Sporulation on the underside of leaves results in the production of sporangia that are dispersed by wind. On arrival on a susceptible host, sporangia germinate in free water on the leaf surface, producing biflagellate zoospores that swim to and encyst on stomata, where they form germ tubes. An appressorium is produced and forms a penetration hypha, which enters the leaf tissue through the stomata. Hyphae grow through the mesophyll and establish haustoria, specialized structures for the transfer of nutrients and signals between host and pathogen. CONTROL Management of downy mildew in Europe requires the use of tolerant cucurbit cultivars in conjunction with fungicide applications. In the USA, an aggressive fungicide programme, with sprays every 5-7 days for cucumber and every 7-10 days for other cucurbits, has been necessary to control outbreaks and to prevent crop loss. USEFUL WEBSITES http://www.daylab.plp.msu.edu/pseudoperonospora-cubensis/ (Day Laboratory website with research advances in downy mildew); http://veggies.msu.edu/ (Hausbeck Laboratory website with downy mildew news for growers); http://cdm.ipmpipe.org/ (Cucurbit downy mildew forecasting homepage); http://ipm.msu.edu/downymildew.htm (Downy mildew information for Michigans vegetable growers).

Advances in Research on Phytophthora capsici on Vegetable Crops in The United States

Since L. H. Leonians first description of Phytophthora capsici as a pathogen of chile pepper in 1922, we have made many advances in our understanding of this pathogens biology, host range, dissemination, and management. P. capsici causes foliar blighting, damping-off, wilting, and root, stem, and fruit rot of susceptible hosts, and economic losses are experienced annually in vegetable crops including cucurbits and peppers. Symptoms of P. capsici infection may manifest as stunting, girdling, or cankers for some cultivars or crops that are less susceptible. P. capsici continues to be a constraint on production, and implementation of an aggressive integrated management scheme can still result in insufficient control when weather is favorable for disease. Management of diseases caused by P. capsici is currently limited by the long-term survival of the pathogen as oospores in the soil, a wide host range, long-distance movement of the pathogen in surface water used for irrigation, the presence of fungicide-resistant pathogen populations, and a lack of commercially acceptable resistant host varieties. P. capsici can infect a wide range of hosts under laboratory and greenhouse conditions including cultivated crops, ornamentals, and native plants belonging to diverse plant families. As our understanding of P. capsici continues to grow, future research should focus on developing novel and effective solutions to manage this pathogen and prevent economic losses due to the diseases it causes.

Investigating the Genetic Structure of Phytophthora capsici Populations

Phytophthora capsici Leonian is a destructive soilborne pathogen that infects economically important solanaceous, cucurbitaceous, fabaceous, and other crops in the United States and worldwide. The objective of this study was to investigate the genetic structure of 255 P. capsici isolates assigned to predefined host, geographical, mefenoxam-sensitivity, and mating-type categories. Isolates from six continents, 21 countries, 19 U.S. states, and 26 host species were genotyped for four mitochondrial and six nuclear loci. Bayesian clustering revealed some population structure by host, geographic origin, and mefenoxam sensitivity, with some clusters occurring more or less frequently in particular categories. Bayesian clustering, split networks, and statistical parsimony genealogies also detected the presence of non-P. capsici individuals in our sample corresponding to P. tropicalis (n = 9) and isolates of a distinct cluster closely related to P. capsici and P. tropicalis (n = 10). Our findings of genetic structuring in P. capsici populations highlight the importance of including isolates from all detected clusters that represent the genetic variation in P. capsici for development of diagnostic tools, fungicides, and host resistance. The population structure detected will also impact the design and interpretation of association studies in P. capsici. This study provides an initial map of global population structure of P. capsici but continued genotyping of isolates will be necessary to expand our knowledge of genetic variation in this important plant pathogen.

The genetic structure of Pseudoperonospora cubensis populations

Pseudoperonospora cubensis is a destructive foliar pathogen of economically important cucurbitaceous crops in the United States and worldwide. In this study, we investigated the genetic structure of 465 P. cubensis isolates from three continents, 13 countries, 19 states of the United States, and five host species using five nuclear and two mitochondrial loci. Bayesian clustering resolved six genetic clusters and suggested some population structure by geographic origin and host, because some clusters occurred more or less frequently in particular categories. All of the genetic clusters were present in the sampling from North America and Europe. Differences in cluster occurrence were observed by country and state. Isolates from cucumber had different cluster composition and lower genetic diversity than isolates from other cucurbits. Because genetic structuring was detected, isolates that represent the genetic variation in P. cubensis should be used when developing diagnostic tools, fungicides, and resistant host varieties. Although this study provides an initial map of global population structure of P. cubensis, future genotyping of isolates could reveal population structure within specific geographic regions, across a wider range of hosts, or during different time points during the growing season.

The Genetic Structure of Pseudoperonospora cubensis Populations

Pseudoperonospora cubensis is a destructive foliar pathogen of economically important cucurbitaceous crops in the United States and worldwide. In this study, we investigated the genetic structure of 465 P. cubensis isolates from three continents, 13 countries, 19 states of the United States, and five host species using five nuclear and two mitochondrial loci. Bayesian clustering resolved six genetic clusters and suggested some population structure by geographic origin and host, because some clusters occurred more or less frequently in particular categories. All of the genetic clusters were present in the sampling from North America and Europe. Differences in cluster occurrence were observed by country and state. Isolates from cucumber had different cluster composition and lower genetic diversity than isolates from other cucurbits. Because genetic structuring was detected, isolates that represent the genetic variation in P. cubensis should be used when developing diagnostic tools, fungicides, and resistant host varieties. Although this study provides an initial map of global population structure of P. cubensis, future genotyping of isolates could reveal population structure within specific geographic regions, across a wider range of hosts, or during different time points during the growing season.

Effects of Temperature, Concentration, Age, and Algaecides on Phytophthora capsici Zoospore Infectivity

Controlled laboratory studies were undertaken to determine the effects of water temperature (2, 9, 12, 19, 22, and 32°C), inoculum concentration (1 × 102, 1 × 103, 5 × 103, 1 × 104, 2 × 104, and 4 × 104 zoospores/ml), and zoospore suspension age (0, 1, 3, and 5 days old) on infection of pickling cucumbers (Cucumis sativus) by Phytophthora capsici. Zoospore motility and mortality in response to commercial algaecides were also investigated. Cucumbers became infected at all temperatures tested, except 2°C, and the highest infection incidence was observed for cucumbers incubated in suspensions held at ≥19°C. Fewer fruit (<40% at ≥19°C, 0% at ≤12°C) became infected when water contained 1 × 102 zoospores/ml. Almost 100% of fruit were infected when water contained ≥5 × 103 zoospores/ml at temperatures ≥12°C. While the incidence of fruit infection declined with the zoospore suspension age, infection still occurred when 5-day-old suspensions were used. Commercial algaecides inhibited zoospore motility and caused significant zoospore mortality in laboratory assays, and show promise for treatment of infested irrigation water. Avoidance of infested irrigation water throughout the growing season is warranted until effective and economically acceptable water treatments are developed for field use.

Relationships Between Airborne Pseudoperonospora cubensis Sporangia, Environmental Conditions, and Cucumber Downy Mildew Severity

Airborne Pseudoperonospora cubensis sporangia were collected 0.5 m above the ground from May to late September or early October 2010 and 2011 in unsprayed cucumber (Cucumis sativus) fields in Benton Harbor and Frankenmuth, MI. Cucumber downy mildew incidence and severity were evaluated weekly within each field from June until September or October. The first airborne sporangium was detected before the crop was planted for each site-year. The greatest numbers of airborne sporangia were detected when moderate to high disease severity (≥5% symptomatic leaf area) was detected within the field. Fewer airborne sporangia were present with low disease severity (<5% symptomatic leaf area), and even fewer were detected prior to planting the cucumber crop. The number of airborne sporangia detected, time post planting, planting number (first versus second versus third planting), temperature, and leaf wetness were positively associated and solar radiation was negatively associated with disease occurrence for at least one site-year. Michigan growers currently use an aggressive, calendar-based fungicide program to manage cucumber downy mildew. Because airborne sporangium concentrations were one of the most important factors identified in this study, the current fungicide recommendation of decreasing the spray interval following disease detection in an area is warranted.

Differences in virulence of Phytophthora capsici isolates from a worldwide collection on host fruits

Phytophthora capsici causes root, crown, and fruit rot of vegetable and tropical hosts. Cucumber, zucchini, tomato, and pepper fruits were inoculated using 6-mm-diameter agar plugs of P. capsici, incubated in clear plastic boxes at room temperature (25 ± 2°C and 100% relative humidity), and virulence was estimated by measuring the lesion diameter, pathogen growth diameter, and pathogen sporulation density three (cucumber, zucchini) or four (tomato, pepper) days later. When isolates were grouped by genetic cluster, significant differences in virulence were observed on cucumber and zucchini, with isolates belonging to genetic cluster five causing larger lesions than isolates from genetic cluster six. On tomato, no significant differences were observed for isolates grouped by genetic cluster, but isolates from vegetable crops were generally more virulent than isolates from tropical hosts. Isolates from fabaceous hosts sporulated better on cucumber fruits than isolates from solanaceous hosts. Isolates from vegetable hosts sporulated better on zucchini than isolates from tropical hosts. No significant differences in lesion diameter were noted on pepper when isolates were grouped by host family of origin or genetic cluster, but differences in pathogen sporulation were apparent by host family. Our findings suggest that isolate characteristics such as host family of origin and genetic cluster membership may be used to guide initial isolate selection for cucurbit fruit resistance screening. Final isolate selection should incorporate the phenotypic and genetic diversity of P. capsici, including isolates with differing virulence to the host organ of interest.

Variation in Phenotypic Characteristics of Phytophthora capsici Isolates from a Worldwide Collection

To determine variation within Phytophthora capsici, 124 P. capsici isolates from 12 countries were characterized for sporangial length and width, pedicle length, oospore diameter, sporangia and chlamydospore production, and growth at 32, 35, and 38°C. Sporangia were 23 to 35 μm wide and 38 to 60 μm long; differences in width and length were noted when isolates were grouped by genetic cluster and continent of origin. Length:breadth ratio (1.34 to 2.07) and pedicle length (20 to 260 μm long) varied widely among isolates; differences were apparent by continent and host family of origin. Oospore diameters varied among isolates (22 to 37 μm), but no differences were noted by isolate genetic cluster, host family of origin, continent of origin, mating type, or sensitivity to mefenoxam. Differences in sporangia production were observed among isolates grouped by continent, and isolates from nonvegetable hosts produced fewer sporangia than isolates from vegetable hosts. When cultures were incubated in a liquid medium, 35 P. capsici isolates formed chlamydospores. Most (122 of 124) of the isolates were able to grow at 35°C, but all of the isolates grew poorly at 38°C. The results of this study indicate substantial variation in morphological and physiological characteristics among P. capsici isolates.